The cornea is a transparent structure composed of three layers: the epithelium, the stroma and the endothelium. To maintain its ransparency the stroma remains in a constant state of dehydration. Consequently, any ion flow disorder through the covering layers can compromise the barrier function and, therefore the corneal homeostasis. Since ionic permeability has a fundamental impact on the passive electrical properties of living tissues, in this work it is proposed and demonstrated a diagnosis method based on tetrapolar impedance measurements performed by electrodes placed on the corneal surface. The contribution of each cornea layer to the total measured impedance has been analysed over a frequency range. Following the obtained guidelines, a flexible probe with integrated electrodes has been developed and manufactured using SU-8 photoresin. The feasibility of the proposed method has been evaluated in vivo by monitoring corneal epithelium wound healing. Obtained impedance measurements have been compared with measurements of permeability to sodium fluorescein from different excised corneas. Successful results demonstrate the feasibility of this novel flexible sensor and its capability to quantify corneal permeability in vivo in a non-invasive way.